Dual Functions of a Rubisco Activase in Metabolic Repair and Recruitment to Carboxysomes

Flecken, Mirkko; Wang, Huping; Popilka, L.; Hartl, F. Ulrich; Bracher, A.; Hayer‐Hartl, Manajit

doi:10.1016/j.cell.2020.09.010

Cited by 31 publications

(22 citation statements)

References 110 publications

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“…All relevant biochemical experiments were replicated two or three times. No statistical methods were used to predetermine sample size, but our sample sizes are similar to those reported in previous publications 16 , 19 . For cryo-EM, data were screened on eight independently prepared samples.…”

Section: Methodsmentioning

confidence: 64%

“…The restructuring of the pre-carboxysome condensate following oxidation may promote the formation of channels around the Rubisco lattice that can be navigated by other carboxysomal proteins and metabolites. This would also facilitate the metabolic repair of Rubisco by Rubisco activase, which possesses SSUL modules for recruitment into the pre-carboxysome matrix 19 .…”

Section: Discussionmentioning

confidence: 99%

“…Implementation of a carboxysome-like CO 2 -concentrating mechanism (CCM) in chloroplasts as a strategy for improving photosynthetic efficiency in crop plants requires a detailed understanding of carboxysome biogenesis 5 – 15 . Recent advances have shown that, early in this process, specialized scaffolding proteins initiate phase separation of Rubisco into a condensate for subsequent encapsulation 16 – 19 . However, the mechanisms underlying the sequestration of CA together with Rubisco are not yet understood.…”

Section: Mainmentioning

confidence: 99%

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Scaffolding protein CcmM directs multiprotein phase separation in β-carboxysome biogenesis

Zang

Wang

Hartl

et al. 2021

Nat Struct Mol Biol

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Carboxysomes in cyanobacteria enclose the enzymes Rubisco and carbonic anhydrase to optimize photosynthetic carbon fixation. Understanding carboxysome assembly has implications in agricultural biotechnology. Here we analyzed the role of the scaffolding protein CcmM of the β-cyanobacterium Synechococcus elongatus PCC 7942 in sequestrating the hexadecameric Rubisco and the tetrameric carbonic anhydrase, CcaA. We find that the trimeric CcmM, consisting of γCAL oligomerization domains and linked small subunit-like (SSUL) modules, plays a central role in mediation of pre-carboxysome condensate formation through multivalent, cooperative interactions. The γCAL domains interact with the C-terminal tails of the CcaA subunits and additionally mediate a head-to-head association of CcmM trimers. Interestingly, SSUL modules, besides their known function in recruiting Rubisco, also participate in intermolecular interactions with the γCAL domains, providing further valency for network formation. Our findings reveal the mechanism by which CcmM functions as a central organizer of the pre-carboxysome multiprotein matrix, concentrating the core components Rubisco and CcaA before β-carboxysome shell formation.

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Section: Methodsmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Mainmentioning

confidence: 99%

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Scaffolding protein CcmM directs multiprotein phase separation in β-carboxysome biogenesis

Zang

Wang

Hartl

et al. 2021

Nat Struct Mol Biol

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“…In the bacterial carboxysomal α-system, we have demonstrated that Rubisco binds the intrinsically disordered proteins CsoS2 (26) and CsoSCA. Additionally, it also binds the Rubisco activase CbbQO, likely via CbbO's von Willebrand factor A domain (36,37) The β-carboxysomal Rubisco binds its interaction partners, the scaffolding protein CcmM (38) (39), and the Rubisco activase Rca (40,41), via a folded domain resembling the small subunit of Rubisco. Despite convergent evolution, a notable similarity between both carboxysome systems is the binding site on Rubisco.…”

Section: Rubisco As An Interaction Hub In Biophysical Ccm'smentioning

confidence: 99%

Discovery of a carbonic anhydrase-Rubisco complex within the alpha-carboxysome

Blikstad

Dugan

Laughlin

et al. 2021

Preprint

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Carboxysomes are proteinaceous organelles that encapsulate key enzymes of CO2 fixation, Rubisco and carbonic anhydrase, and are the centerpiece of the bacterial CO2 concentrating mechanism (CCM). In the CCM, actively accumulated cytosolic bicarbonate diffuses into the carboxysome and is converted to CO2 by carbonic anhydrase, producing a high CO2 concentration near Rubisco and ensuring efficient carboxylation. Self-assembly of the α-carboxysome is orchestrated by the intrinsically disordered scaffolding protein, CsoS2, which interacts with both Rubisco and carboxysomal shell proteins, but it is unknown how CsoSCA, the carbonic anhydrase, is incorporated into the α-carboxysome. Here, we present the structural basis of carbonic anhydrase encapsulation into α-carboxysomes from Halothiobacillus neapolitanus. We find that CsoSCA interacts directly with Rubisco via an intrinsically disordered N-terminal domain. A 1.98 Å single-particle cryo-electron microscopy structure of Rubisco in complex with this peptide reveals that CsoSCA binding is predominantly mediated by a network of hydrogen bonds. CsoSCAs binding site overlaps with that of CsoS2 but the two proteins utilize substantially different motifs and modes of binding, revealing a plasticity of the Rubisco binding site. Our results advance the understanding of biogenesis of carboxysomes and highlights the importance of Rubisco, not only as an enzyme, but also as a hub protein central for assembling supercomplexes.

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“…With regards to Form IB Rubisco, it has very recently been shown that the Rca-like protein found in some cyanobacterial clades (38,39) is a functional Rubisco activase in Nostoc sp. PCC7120 (40).…”

Section: Introductionmentioning

confidence: 99%

The CbbQO-type rubisco activases encoded in carboxysome gene clusters can activate carboxysomal form IA rubiscos

Tsai¹,

Liew²,

Guo

et al. 2022

Journal of Biological Chemistry

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Dual Functions of a Rubisco Activase in Metabolic Repair and Recruitment to Carboxysomes

Cited by 31 publications

References 110 publications

Scaffolding protein CcmM directs multiprotein phase separation in β-carboxysome biogenesis

Scaffolding protein CcmM directs multiprotein phase separation in β-carboxysome biogenesis

Discovery of a carbonic anhydrase-Rubisco complex within the alpha-carboxysome

The CbbQO-type rubisco activases encoded in carboxysome gene clusters can activate carboxysomal form IA rubiscos

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